一种使用单克隆抗体的人绒毛膜促性腺激素新型放射免疫测定法。

A new radioimmunoassay for human chorionic gonadotropin using monoclonal antibody.

作者信息

Katoh S, Hattori S, Furuhashi Y, Mano H, Goto S, Tomoda Y

出版信息

Endocrinol Jpn. 1986 Oct;33(5):691-700. doi: 10.1507/endocrj1954.33.691.

DOI:10.1507/endocrj1954.33.691

Abstract

A new radioimmunoassay (RIA) for human Chorionic Gonadotropin (hCG) was developed using murine monoclonal antibody to the beta-subunit of hCG (beta-hCG). The IgG fraction of the monoclonal antibody which did not react with 125I-beta-hCG was purified from hybridoma ascites, and covalently coupled to Sepharose 4B. This solid-phase antibody was incubated with standard hCG or serum sampled for 48 hours. The reaction medium was then removed by centrifugation and 125I-beta-hCG and anti-beta-hCG rabbit polyclonal antibody were added to the precipitate. The alcohol precipitation method was used for separating "bound" and "free" forms in the second reaction. The sensitivity for hCG in this assay system was 0.5 mIU/ml serum and the cross-reactivity with human Luteinizing Hormone (hLH) was 0.4%. This assay system was shown to be clinically applicable. Serial serum samples from two patients with trophoblastic disease were assayed and minute amounts of hCG, which could not be determined by conventional assay methods, could be assayed by this new RIA.

摘要

利用针对人绒毛膜促性腺激素（hCG）β亚基（β-hCG）的鼠单克隆抗体，开发了一种新的人绒毛膜促性腺激素放射免疫测定法（RIA）。从杂交瘤腹水中纯化出不与125I-β-hCG发生反应的单克隆抗体的IgG部分，并将其与琼脂糖4B共价偶联。将这种固相抗体与标准hCG或采集48小时的血清样本一起孵育。然后通过离心去除反应介质，并向沉淀物中加入125I-β-hCG和抗β-hCG兔多克隆抗体。在第二个反应中采用乙醇沉淀法分离“结合”和“游离”形式。该测定系统对血清中hCG的灵敏度为0.5 mIU/ml，与人促黄体生成素（hLH）的交叉反应率为0.4%。结果表明该测定系统具有临床应用价值。对两名滋养层疾病患者的系列血清样本进行了检测，用这种新的放射免疫测定法能够检测出微量hCG，而用传统检测方法无法检测到这些微量hCG。